Extraction of Total RNA from Tissues and Cultured Cells

  • Sandeep Raha
  • Mingfu Ling
  • Frank Merante
Part of the Springer Protocols Handbooks book series (SPH)


The rsolatron of intact, high quality, total cellular RNA is often the starting point for many molecular biologrcal procedures (Fig. 1) There are numerous general methods for the rsolation of total cellular RNA (1, 2, 3, 4, 5, 6, 7, 8, 9). There are also many specialrzed methods for the rsolatron of RNA from specific tissues (8, 9, 10), various cell types (11), and subcellular organelles (7,12,13) In addmon, a number of methods describe the srmultaneous rsolatton of RNA and DNA (14, 15, 16, 17). Generally, the rationale for any tsolation procedure is to solubihze cellular components and simultaneously mactrvate mtracellular RNases while mamtaming brologrcally active RNA Therefore, the goal is to acquire purrfied cellular RNA in an intact form that can be a substrate for further mampulatrons, such as in vitro translation, RNase protectton, reverse transcrrptron, and Northern-blot analysis.


RNase Activity Subcellular Organelle Polymerase Chain Reaction Procedure Subsequent Polymerase Chain Reaction Minimal Sample Handling 
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  1. 1.
    Chirgwin, J. M., Przybyla, A. E., MacDonald, R. J., and Rutter, W. J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18,5394–5399.CrossRefGoogle Scholar
  2. 2.
    Auffray, C. and Rougeon, F. (1980) Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur. J. Biochem. 197,303–414.Google Scholar
  3. 3.
    Chomczynski, P. and Sacchi, N. (1987) Single step method of RNA extraction by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162, 156–159.PubMedCrossRefGoogle Scholar
  4. 4.
    Emmett, M. and Petrack, B. (1988) Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells. Anal. Biochem. 173,93–95.CrossRefGoogle Scholar
  5. 5.
    Gough, N in (1988) Rapid and quantltatlve preparation of cytoplasmic RNA from small numbers of cells Anal Biochem 173,93–95PubMedCrossRefGoogle Scholar
  6. 6.
    Meler, R (1988) A universal and efficient protocol for the isolation of RNA from tissues and cultured cells Nucleic Acids Res 16,23–40Google Scholar
  7. 7.
    Wilkinson, M (1988) A rapid and convenient method for lsolatlon of nuclear, cytoplasmlc and total cellular RNA Nucleic Acids Res 16, 10–934Google Scholar
  8. 8.
    Nemeth, G G, Heydemann, A, and Bolander, M E (1989) Isolation and analysis of ribonucleic acids from skeletal tissues Anal Biochem 183,301–304PubMedCrossRefGoogle Scholar
  9. 9.
    Tavangar, K, Hoffman, A R, and Kraemer, F B (1990) A micromethod for the isolation of total RNA from adipose tissue Anal Biochem 186,60–63PubMedCrossRefGoogle Scholar
  10. 10.
    Monstem, H-J, Nylander, A-G, and Chen, D (1995) RNA extraction from gastrointestinal tract and pancreas by a modified Chomczynsla and Sacchi method Biotechniques 10,341–344Google Scholar
  11. 11.
    Vauti, F and Sless, W (1993) Simple method of RNA isolation from human leucocytic cell lines Nucleic Acids Res 23,4852,4853Google Scholar
  12. 12.
    Wilkinson, M (1988) RNA lsolatlon a mini-prep method Nucleic Acids Res 16, 10,933PubMedCrossRefGoogle Scholar
  13. 13.
    Hatch, C L and Bonner, W in (1987) Direct analysis of RNA in whole cell and cytoplasmic extracts by gel electrophoresls Anal Biochem 162,283–290PubMedCrossRefGoogle Scholar
  14. 14.
    Raha, S, Merante, F, Proteau, G, and Reed, J K (1990) Simultaneous isolation of total cellular RNA and DNA from tissue culture cells using phenol and lithum chloride Gene Anal Tech 7, 173–177CrossRefGoogle Scholar
  15. 15.
    Merante, F, Raha, S, Reed, J K, and Proteau, G (1994) The simultaneous isolation of RNA and DNA from tissues and cultured cells Methods Mol Biol 32, 113–120Google Scholar
  16. 16.
    Kotlinska, J and Wojcierowski, (1995) RNA isolation from solid tumor tissue Anal Biochem 228, 170–172CrossRefGoogle Scholar
  17. 17.
    Coombs, L in, Pigott, D, Proctor, A, Edymann, M, Denner, J, and Knowles, M A (1990) Simultaneous lsolatlon of DNA, RNA and antigenic protein exhlblting kmase actlvlty from small tour samples using guanidine isothiocyanate Anal Biochem 188,338–343PubMedCrossRefGoogle Scholar
  18. 18.
    Ogretmen, B and Saffa, A R (1995) Mini-preparation of total RNA for RT-PCR from cultured human cells Biotechniques 19,374–376PubMedGoogle Scholar
  19. 19.
    Chattopadhyay, N, Kher, R, and Godbole, M (1993) Inexpensive SDS/phenol method from RNA extraction from tissues Biotechniques 15,26Google Scholar
  20. 20.
    Dhale, C E and MacFarlane, D E (1993) Isolation of RNA from cells in culture using Catrimox-14 cationic surfactant Biotechniques 15, 1102–1105Google Scholar
  21. 21.
    Chan, V T-W, Flemming, K A, and McGee, J O’D (1988) Simultaneous extraction from clinical biopsies of high-molecular-weight DNA and RNA comparative charactenzation by biotinylated and 32P-labeled probes on Southern and Northern blots Anal Biochem 168,16–26PubMedCrossRefGoogle Scholar
  22. 22.
    Zarlenga, D S and Gamble, H R (1987) Simultaneous isolatlon of preparative amounts of RNA and DNA from Trichwella spiralls by cesmm trlbluoroacetate isopycnic centrifugation Anal Biochem 162, 569–574PubMedCrossRefGoogle Scholar
  23. 23.
    Mirkes, P E (1985) Simultaneous banding of rat embryo DNA, RNA and protein in cesmm trifluoroacetate gradients Anal Biochem 148, 376–383PubMedCrossRefGoogle Scholar
  24. 24.
    Karlmsey, J, Stamatoyannopoulos, G, and Enver, T (1989) Simultaneous purification of DNA and RNA from small numbers of eukaryotic cells Anal Biochem 180,303–306CrossRefGoogle Scholar
  25. 25.
    Slebert, P D and Chenchik, A (1993) Modified acid guanidium thlocyanate-phenolchloroform RNA extraction method which greatly reduces DNA contamination Nucleic Acids Res 23,2019,2020Google Scholar
  26. 26.
    Kneg, P, Amtmann, E, and Sauer, G (1983) The simultaneous extraction of high-molecular-weight DNA and RNA from sohd tumors Anal Biochem 134,288–294CrossRefGoogle Scholar
  27. 27.
    Kingston, R E, Chomczynskl, P, and Sacchi, N (1988) Single-step RNA isolation from cultured cells or tissues, in Current Protocols in Molecular Biology (Ausubel, S in, Brent, R, Kingston, R E, Moore, D D, Seldman, J G, Smith, J A, Struhl, K, eds ), ngreen, Brooklyn, NY, pp 424–428Google Scholar
  28. 28.
    Kormanec, J and Farkasovsky, M (1994) Isolation of total RNA from yeast and bacteria and detectlon of rRNA in Northern blots Biotechniques 71, 838–842Google Scholar
  29. 29.
    Bugos, R C, Chlang, V L, Zhang, X H, Campbell, E R, Pohda, G K, and Campbell, W H (1995) RNA isolation from plant tissues recalcitrant to extraction in guanidine Biotechniques 19, 734–737PubMedGoogle Scholar
  30. 30.
    Fourney, R in, Miyakoshi, J, Day, R S, and Paterson, M C (1988) Northern blotting efficient RNA staining and transfer Focus 10,5–7Google Scholar
  31. 31.
    Monstem, H, Nylander, G, and Chen, D (1995) RNA extractlon from gastromtestmal tract and pancreas by a modified Chomczynskl and Sacchi method Biotech 19,340–343Google Scholar
  32. 32.
    Noonberg, S B, Scott, G K, and Benz, C C (1995) Effect of pH on RNA degradation during guamdmmm extractlon Biotechniques 19,731–733PubMedGoogle Scholar

Copyright information

© Humana Press Inc , Totowa, NJ 1998

Authors and Affiliations

  • Sandeep Raha
    • 1
  • Mingfu Ling
    • 1
  • Frank Merante
    • 1
  1. 1.Department of GeneticsHospital for Sick ChildrenCanada

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